Institute of Organic Chemistry, University of Münster, Corrensstrasse 40, D-48149 Münster, Germany.
Institute of Medical Biochemistry, Center for Molecular Biology of Inflammation, University of Münster, Von-Esmarch-Strasse 56, D-48149 Münster, Germany.
Cell Chem Biol. 2018 Aug 16;25(8):952-961.e12. doi: 10.1016/j.chembiol.2018.04.015. Epub 2018 May 24.
Cholesterol is an essential component of most biological membranes and serves important functions in controlling membrane integrity, organization, and signaling. However, probes to follow the dynamic distribution of cholesterol in live cells are scarce and so far show only limited applicability. Herein, we addressed this problem by synthesizing and characterizing a class of versatile and clickable cholesterol-based imidazolium salts. We show that these cholesterol analogs faithfully mimic the biophysical properties of natural cholesterol in phospholipid mono- and bilayers, and that they integrate into the plasma membrane of cultured and primary human cells. The membrane-incorporated cholesterol analogs can be specifically labeled by click chemistry and visualized in live-cell imaging experiments that show a distribution and behavior comparable with that of endogenous membrane cholesterol. These results indicate that the cholesterol analogs can be used to reveal the dynamic distribution of cholesterol in live cells.
胆固醇是大多数生物膜的重要组成部分，在控制膜的完整性、组织和信号传递方面发挥着重要作用。然而，用于跟踪胆固醇在活细胞中动态分布的探针却很少，到目前为止，这些探针的适用性非常有限。为了解决这个问题，我们合成并表征了一类多功能的可点击的基于胆固醇的咪唑鎓盐。我们发现，这些胆固醇类似物能够真实地模拟天然胆固醇在磷脂单分子层和双分子层中的生物物理特性，并且能够整合到培养的和原代人细胞的质膜中。整合到膜中的胆固醇类似物可以通过点击化学进行特异性标记，并在活细胞成像实验中可视化，其分布和行为与内源性膜胆固醇相似。这些结果表明，胆固醇类似物可用于揭示活细胞中胆固醇的动态分布。
